Protoplasts from pectinolytic fungi: isolation, regeneration and pectinolytic enzyme production

Abstract

S. Solís, M.E. FLORES AND C. HUITRON. 1996. Protoplast release in pectinolytic strain mutants of Aspergillus sp. CH‐Y‐1043 (A13) and Aspergillus flavipes ATCC‐16795 (F7) is described. Optimum yield of protoplasts A13 was obtained in a lapse of 1 h when commercially lytic enzymes of Trichoderma harzanium (2 mg ml−1) were added in 0.05 mol 1−1 citrate‐phosphate buffer pH 5.0 containing 0.7 mol 1−1 KCl and 10 mg ml−1 BSA. Best results in F7 were obtained when the protoplasting system of A13 was supplemented with 10 mg ml−1Aureobasidium sp. lytic enzymes. Isolated protoplasts in A13 and F7 were capable of a high regeneration frequency of 87% and 53% when 0.7 mol 1−1 KCl and sorbitol were used as osmotic stabilizers. Endo‐P, Exo‐P and pectin lyase production were not modified during the process of regeneration.